Generation of human CAR-T cells with tissue-resident memory features for improved treatment of hepatocellular carcinoma

Primary supervisor: Anna Schurich, King’s College London

Secondary supervisor: Laura Pallett, UCL

Project

Project Background & Description:

Liver cancer (hepatocellular carcinoma [HCC]) remains the third leading cause of cancer-related deaths worldwide, with the incidence rising [1]. Late diagnosis, the limits of surgical intervention and resistance to chemotherapy mean that few patients are cured of their disease [2]. Over recent years a shift to immunotherapy has revolutionised treatment options for HCC, however despite the combination of checkpoint inhibitors becoming first-line therapy for unresectable disease, only 30% of patients respond [2].

CAR T-cell therapy, utilising genetic engineering of circulating, non-antigen specific T-cells, has great potential for HCC [3]. To realise treatment success equal to that achieved in blood-cancer, it is crucial to understand and overcome the restrictions imposed on CAR T-cells by the tumour microenvironment (TME). We hypothesise that to achieve effective and durable treatment for HCC we need to impart CAR T-cells with the appropriate characteristics to function both within the immunoregulatory liver environment and the HCC-TME. Our work has previously characterised tissue-resident CD8+T-cells (CD8+TRM) in the liver, confirming these cells to be highly-specialised effectors [4], with other groups confirming a correlation of CD8+TRM frequency with increased relapse-free survival in HCC [5].
Our key research goal is to define the characteristics of CAR T-cells induced in vitro to have the transcriptional, phenotypic and functional profile of CD8+TRM – “CAR-TRM”. Specifically investigating their potential to mediate improved anti-tumour responses compared to conventional CAR T-cells. We reason our method could be utilised in addition to other genetic manipulations/strategies in development to specifically direct CAR T-cells to the HCC and enhance their effector function.

During your PhD, you will work closely with the Schurich-lab and the Pallett-lab using state-of-the-art methods and techniques. Drawing on our strong expertise in T-cell biology and immunotherapeutics this project will investigate the mechanisms critical for improved CAR-T-cell homing, long-term retention, and enhanced survival that would in turn contribute to an increased ability to kill tumour cells within the challenging liver microenvironment. In the first phase of your PhD, you will spend some time at Leucid Bio (King’s College London) to acquire expertise in the process of genetically engineering CAR T-cells. This will be followed by the detailed characterisation of the phenotypic, metabolic and functional features of HCC-specific in vitro induced CAR-TRM using technologies such as flow-cytometry, high-dimensional imaging (MACSima), metabolomics and metabolic-flux analysis. In the second phase you will investigate the function of induced CAR-TRM in CAR T- cell/tumour cell co-culture models under conditions of metabolic stress. You will undertake a research-rotation with Dr Sinclair (University of Dundee) receiving training in the assessment of nutrient uptake using click-chemistry. Finally, you will gain experience in mass-cytometry in the lab of Dr Tape (Cancer Institute; UCL) unravelling the cellular communication networks involved in the novel characteristics of CAR-TRM. Here you will also gain expertise in large data-set analysis.

Candidate background

We are particularly interested in receiving applications from candidates with a background in immunology, cancer biology or related subjects with a strong interest in translating fundamental research into novel therapeutics for HCC.

Potential Research Placements

1. John Maher, CAR-mechanics research group, King’s College London
2. Linda Sinclair, University of Dundee
3. Chris Tape, UCL Cancer Institute

References

1. Kumari, R., Sahu, M.K., Tripathy, A., Uthansingh, K., and Behera, M. (2018). Hepatocellular carcinoma treatment: hurdles, advances and prospects. Hepatic Oncol 5, HEP08. 10.2217/hep- 2018-0002.
2. Llovet, J.M., Castet, F., Heikenwalder, M., Maini, M.K., Mazzaferro, V., Pinato, D.J., Pikarsky, E., Zhu, A.X., and Finn, R.S. (2022). Immunotherapies for hepatocellular carcinoma. Nat Rev Clin Oncol 19, 151-172. 10.1038/s41571-021-00573-2.
3. Guo, J., and Tang, Q. (2021). Recent updates on chimeric antigen receptor T cell therapy for hepatocellular carcinoma. Cancer Gene Ther 28, 1075-1087. 10.1038/s41417-020-00259-4.
4. Pallett, L.J., Davies, J., Colbeck, E.J., Robertson, F., Hansi, N., Easom, N.J.W., Burton, A.R., Stegmann, K.A., Schurich, A., Swadling, L., et al. (2017). IL-2(high) tissue-resident T cells in the human liver: Sentinels for hepatotropic infection. J. Exp. Med. 214, 1567-1580. 10.1084/jem.20162115.
5. Cheng, Y., Gunasegaran, B., Singh, H.D., Dutertre, C.-A., Loh, C.Y., Lim, J.Q., Crawford, J.C., Lee, H.K., Zhang, X., Lee, B., et al. (2021). Non-terminally exhausted tumor-resident memory HBV- specific T cell responses correlate with relapse-free survival in hepatocellular carcinoma. Immunity 54, 1825-1840.e7. 10.1016/j.immuni.2021.06.013.